巢式实时 SYBR 绿 RT-PCR 检测法诊断巴尔马森林病毒感染。
Diagnosis of Barmah Forest virus infection by a nested real-time SYBR green RT-PCR assay.
机构信息
Centre for Infectious Diseases and Microbiology Laboratory Services, Westmead, NSW, Australia.
出版信息
PLoS One. 2013 Jul 23;8(7):e65197. doi: 10.1371/journal.pone.0065197. Print 2013.
Barmah Forest virus (BFV) is a mosquito borne (+) ssRNA alphavirus found only in Australia. It causes rash, myalgia and arthralgia in humans and is usually diagnosed serologically. We developed a real-time PCR assay to detect BFV in an effort to improve diagnosis early in the course of infection. The limit of detection was 16 genome equivalents with a specificity of 100%. Fifty five serum samples from BFV-infected patients were tested by the PCR. 52 of 53 antibody-positive samples were PCR negative. Two culture-positive (neutralizing antibody negative) samples were positive on first round PCR, while one sample (IgM and neutralizing antibody strongly positive, IgG negative) was positive on second round PCR, suggesting that viral RNA is detectable and transiently present in early infection. PCR can provide results faster than culture, is capable of high throughput and by sequencing the PCR product strain variants can be characterized.
巴尔马森林病毒（BFV）是一种仅在澳大利亚发现的蚊虫传播（+）ssRNA 甲病毒。它会导致人类出现皮疹、肌肉痛和关节痛，通常通过血清学诊断。我们开发了一种实时 PCR 检测方法，以提高感染早期的诊断能力。该检测方法的检测限为 16 个基因组当量，特异性为 100%。用该 PCR 检测了 55 份 BFV 感染患者的血清样本。53 份抗体阳性样本中，52 份 PCR 检测为阴性。2 份培养阳性（中和抗体阴性）的样本在第一轮 PCR 中呈阳性，而 1 份样本（IgM 和中和抗体强阳性，IgG 阴性）在第二轮 PCR 中呈阳性，提示在早期感染中可检测到并短暂存在病毒 RNA。PCR 比培养更快提供结果，能够高通量检测，并且通过对 PCR 产物进行测序可以对病毒株进行特征分析。
Zotero 插件